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Characterizing the Non-Normal Distribution of Flow Cytometry Measurements from Transiently Expressed Constructs in Mammalian Cells

Peter F. McLean, Christina D. Smolke, Marc Salit
doi: https://doi.org/10.1101/057950
Peter F. McLean
1Joint Initiative for Metrology in Biology, National Institute of Standards and Technology, Stanford, CA
2Department of Bioengineering, Stanford University, CA
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Christina D. Smolke
2Department of Bioengineering, Stanford University, CA
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Marc Salit
1Joint Initiative for Metrology in Biology, National Institute of Standards and Technology, Stanford, CA
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Abstract

In mammalian cells, transient gene expression (TGE) is a rapid, minimal-investment alternative to single-copy integrations for testing of transgenic constructs. However, transient gene expression, as measured by flow cytometry with a fluorescent reporter, typically displays a broad, asymmetric distribution with a left-tail that is convolved with background signal. Common approaches for deriving a summary statistic for transiently expressed gene products impose a normal distribution on gated or ungated data. Summary statistics derived from these models are heavily biased by experimental conditions and instrument settings that are difficult to replicate and insufficient to accurately describe the underlying data. Here, we present a convolved gamma distribution as a superior model for TGE datasets. The 4-6 parameters of this model are sufficient to accurately describe the entire, ungated distribution of transiently transfected HEK cells expressing monomeric fluorescent proteins, that operates consistently across a range of transfection conditions and instrument settings. Based on these observations, a convolved gamma model of TGE distributions has the potential to significantly improve the accuracy and reproducibility of genetic device characterization in mammalian cells.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted September 30, 2016.
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Characterizing the Non-Normal Distribution of Flow Cytometry Measurements from Transiently Expressed Constructs in Mammalian Cells
Peter F. McLean, Christina D. Smolke, Marc Salit
bioRxiv 057950; doi: https://doi.org/10.1101/057950
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Characterizing the Non-Normal Distribution of Flow Cytometry Measurements from Transiently Expressed Constructs in Mammalian Cells
Peter F. McLean, Christina D. Smolke, Marc Salit
bioRxiv 057950; doi: https://doi.org/10.1101/057950

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